title: Elucidation of carbohydrate molecular interaction mechanism of recombinant and native ArtinM.
abstract: The quartz crystal microbalance (QCM) technique has been applied for monitoring the biorecognition of ArtinM lectins at low horseradish peroxidase glycoprotein (HRP) concentrations, using a simple kinetic model based on Langmuir isotherm in previous work.18 The latter approach was consistent with the data at dilute conditions but it fails to explain the small differences existing in the jArtinM and rArtinM due to ligand binding concentration limit. Here we extend this analysis to differentiate sugar-binding event of recombinant (rArtinM) and native (jArtinM) ArtinM lectins beyond dilute conditions. Equivalently, functionalized quartz crystal microbalance with dissipation monitoring (QCM-D) was used as real-time label-free technique but structural-dependent kinetic features of the interaction were detailed by using combined analysis of mass and dissipation factor variation. The stated kinetic model not only was able to predict the diluted conditions but also allowed to differentiate ArtinM avidities. For instance, it was found that rArtinM avidity is higher than jArtinM avidity whereas their conformational flexibility is lower. Additionally, it was possible to monitor the hydration shell of the binding complex with ArtinM lectins under dynamic conditions. Such information is key in understanding and differentiating protein binding avidity, biological functionality, and kinetics.
authors.affiliation: Institute of Molecular Recognition and Technological Development, Department of Chemistry, Universitat Politècnica de València, Camino de Vera s/n, 46022 Valencia, Spain. giroda@qim.upv.es
authors.forename: David
authors.initials: D
authors.lastname: Giménez-Romero
authors.forename: Paulo R
authors.initials: PR
authors.lastname: Bueno
authors.forename: Naira C
authors.initials: NC
authors.lastname: Pesquero
authors.forename: Isidro S
authors.initials: IS
authors.lastname: Monzó
authors.forename: Rosa
authors.initials: R
authors.lastname: Puchades
authors.forename: Ángel
authors.initials: Á
authors.lastname: Maquieira
mesh_terms: D001665:Binding Sites; D006023:Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D037241:Mannose-Binding Lectins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D037121:Plant Lectins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D058525:Quartz Crystal Microbalance Techniques; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics
chemical_list: D006023:Glycoproteins; D037241:Mannose-Binding Lectins; D037121:Plant Lectins; D011994:Recombinant Proteins; C090683:artocarpin lectin